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Differential Expression of N-myc in Phenotypically Distinct Subclones of a Human Neuroblastoma Cell Line¹

Department of Anatomy and Cell Biology, University of Cincinnati, Cincinnati, Ohio 45267-0521 [J. F., L. M. P.]; Department of Pediatrics, Northwestern University Medical Center Chicago, Illinois 60614 [S. L. C., H. R. S., D. C.]; and Department of Radiation and Cellular Oncology, University of Chicago Medical Center, Chicago, Illinois 60637 [J. C., K. L. M.]

Neuroblastomas are malignant childhood neoplasms that arise from derivatives of the neural crest. We report the characterization of a new neuroblastoma cell line, designated NBL-W, derived from the primary tumor of a patient with stage IVS disease (S. L. Cohn, C. V. Herst, H. S. Maurer, and S. T. Rosen, J. Clin. Oncol., 5: 1441–1444, 1987) according to the criteria of Evans [A. E. Evans, G. J. D'Angio, and J. Randolf, Cancer (Phila.), 27: 374–378, 1971]. Neurite-bearing (N) and substrate-adherent (S) cell lines have been subcloned from the parent line. N and S cells can interconvert, and both cell types label with the neural crest cell surface marker antibody, HNK-1. Cells in the subcloned lines and in the parent line have been shown by Southern blot analysis to contain 100 copies of the N-myc gene. Cytogenetic analysis shows a homogeneously staining region present on chromosome 19. Although these subclones are of identical genotype, the S cells express lower amounts of N-myc mRNA and protein as compared to the N cells. N cells express several neuronal proteins including the neurotransmitter-processing enzymes tyrosine hydroxylase and dopamine ß-hydroxylase, the neuronal intermediate filament proteins peripherin and NF66/-internexin, and the neural cell adhesion molecule. S cells generally lack neuronal markers but express the mesenchymal intermediate filament protein vimentin, and a small subset of the S cells express glial fibrillary acidic protein. Some S cells were labeled weakly with neural cell adhesion molecule antibody; others were negative. S cells did not express the glial marker S-100 or a melanocyte marker, tyrosinase. Thus, S cells express the neural crest marker HNK-1 but do not express a set of antigens characteristic of any known cell type derived from the neural crest. These results are consistent with the suggestion that differential N-myc expression may be involved in the interconversion of N and S cells but indicate that the S cell phenotype need not represent a highly differentiated neural crest derivative.

¹ Supported in part by The Deborah Goldfine Reva Smilgoff Memorial Fund for Cancer Research (S. L. C.), The Shellie Harris Memorial Fund (S. L. C.), the Boothroyd Foundation (L. M. P.), and NIH Grants CA51061 (S. L. C.) and HD25752 (L. M. P.).

² To whom requests for reprints should be sent, at University of Cincinnati, Department of Anatomy and Cell Biology, 231 Bethesda Avenue, (ML 521), Cincinnati, OH 45267-0521.

Received 5/17/91. Accepted 9/19/91.